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Muscle-specific receptor tyrosine kinase (MuSK) is an essential molecule in the NMJ. MUSK exon 10 encodes an important part of the frizzled-like cysteine-rich domain, which is necessary for Wnt-mediated acetylcholine receptors clustering at NMJ. MUSK exon 10 is alternatively spliced in humans but not in mice. We reported that humans acquired a unique exonic splicing silencer in exon 10 compared to mice, which promotes exon skipping coordinated by hnRNP C, YB-1, and hnRNP L. Here, we have dissected the underlying mechanisms of exon inclusion. We precisely characterized the exonic splicing enhancer (ESE) elements and determined the functional motifs. We demonstrated that SRSF6 and SRSF1 coordinately enhance exon inclusion through multiple functional motifs in the ESE. Remarkably, SRSF6 exerts a stronger effect than SRSF1, and SRSF6 alone can compensate the function of SRSF1. Interestingly, differentiated muscle reduces the expression of splicing suppressors, rather than enhancers, to generate a functional Wnt-sensitive MuSK isoform to promote neuromuscular signal transmission. Finally, we developed splice-switching antisense oligonucleotides, which could be used to selectively modulate the expression of MUSK isoforms toward a beneficial outcome for therapeutic intervention.<\/jats:p>","DOI":"10.1093\/narmme\/ugaf007","type":"journal-article","created":{"date-parts":[[2025,3,20]],"date-time":"2025-03-20T02:57:36Z","timestamp":1742439456000},"source":"Crossref","is-referenced-by-count":6,"title":["SRSF6 and SRSF1 coordinately enhance the inclusion of human <i>MUSK<\/i> exon 10 to generate a Wnt-sensitive MuSK isoform"],"prefix":"10.1093","volume":"2","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1130-307X","authenticated-orcid":false,"given":"Farhana","family":"Nasrin","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences , Little Rock, AR 72205 ,","place":["United States"]},{"name":"Division of Neurogenetics, Center for Neurological Diseases and Cancer, Nagoya University Graduate School of Medicine , Nagoya, 4668550 Aichi ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7427-0092","authenticated-orcid":false,"given":"Preeti","family":"Nagar","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences , Little Rock, AR 72205 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3407-2024","authenticated-orcid":false,"given":"Md\u00a0Rafikul","family":"Islam","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences , Little Rock, AR 72205 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-8634-191X","authenticated-orcid":false,"given":"Shabiha\u00a0Afroj","family":"Heeamoni","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences , Little Rock, AR 72205 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-9290-7833","authenticated-orcid":false,"given":"Md\u00a0Mahbub","family":"Hasan","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences , Little Rock, AR 72205 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1529-2750","authenticated-orcid":false,"given":"Kinji","family":"Ohno","sequence":"additional","affiliation":[{"name":"Division of Neurogenetics, Center for Neurological Diseases and Cancer, Nagoya University Graduate School of Medicine , Nagoya, 4668550 Aichi ,","place":["Japan"]},{"name":"Graduate School of Nutritional Sciences, Nagoya University of Arts and Sciences , Nisshin, 4700196 Aichi ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1553-8316","authenticated-orcid":false,"given":"Mohammad\u00a0Alinoor","family":"Rahman","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences , Little Rock, AR 72205 ,","place":["United States"]},{"name":"Winthrop P. Rockefeller Cancer Institute, University of Arkansas for Medical Sciences , Little Rock, AR 72205 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2025,3,19]]},"reference":[{"key":"2025033000432636100_B1","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1016\/j.neuron.2006.09.017","article-title":"Splicing regulation in neurologic disease","volume":"52","author":"Licatalosi","year":"2006","journal-title":"Neuron"},{"key":"2025033000432636100_B2","doi-asserted-by":"publisher","first-page":"777","DOI":"10.1016\/j.cell.2009.02.011","article-title":"RNA and disease","volume":"136","author":"Cooper","year":"2009","journal-title":"Cell"},{"key":"2025033000432636100_B3","doi-asserted-by":"publisher","first-page":"1413","DOI":"10.1038\/ng.259","article-title":"Deep surveying of alternative splicing complexity in the human transcriptome by high-throughput 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